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Toxicological information

Repeated dose toxicity: oral

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Administrative data

Endpoint:
chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Not specified
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study not performed to any reported guidance.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1968
Report date:
1968

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
other: Not specified
GLP compliance:
no
Limit test:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
3,3',3'',5,5',5''-hexa-tert-butyl-α,α',α''-(mesitylene-2,4,6-triyl)tri-p-cresol
EC Number:
216-971-0
EC Name:
3,3',3'',5,5',5''-hexa-tert-butyl-α,α',α''-(mesitylene-2,4,6-triyl)tri-p-cresol
Cas Number:
1709-70-2
Molecular formula:
C54H78O3
IUPAC Name:
3,3',3'',5,5',5''-hexa-tert-butyl-α,α',α''-(mesitylene-2,4,6-triyl)tri-p-cresol
Details on test material:
A 100 kg sample of lonox 330, batch number 89/5/6232, purity 98% was supplied by Shell Chemical Company, Carrington.

Test animals

Species:
dog
Strain:
Beagle
Sex:
male/female
Details on test animals or test system and environmental conditions:
26 Beagle hounds, aged about five months and bred at this laboratory were used. The dogs were inoculated at three months with a vaccine consisting of living, modified, tissue culture adapted distemper virus, inactivated canine hepatitis virus and inactivated leptospira bacteria. They were also treated for nematode parasites with piperazine adipate.

Administration / exposure

Route of administration:
oral: feed
Vehicle:
not specified
Details on oral exposure:
Not specified
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
Not specified
Duration of treatment / exposure:
2 years
Frequency of treatment:
Daily
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 2000, 10000 ppm
Basis:
nominal in diet
No. of animals per sex per dose:
0 - 5 males & 5 females
2000 - 4 males & 4 females
10000 - 4 males & 4 females
Control animals:
yes, plain diet
Details on study design:
Groups of four males and four females, housed individually, received diets containing 2,000 and 10,000 ppm Ionox 330, an intake of 1 ppm in the dogs used for this experiment being equivalent to 0.05 mg/kg/day.
Control groups, five males and five females, received untreated powdered diet (Powdered 'More-Meat' Dog Food. Quaker Oats Ltd., Southall, Middlesex). Dogs within each of the available litters were allocated at random to the various treatments. Here this allocation was not possible animals of a similar age and weight were used.
Positive control:
Not specified

Examinations

Observations and examinations performed and frequency:
Daily records of general health and behaviour were made and body weights were recorded weekly. Blood samples were taken prior to the experiment. After six and 12 weeks exposure and thereafter at three monthly intervals. Haematological examinations carried out on all blood samples comprised haemoglobin and packed cell volume determinations and erythrocyte, leucocyte and di-ferential leucocyte counts. Clinical chemical determinations were made of serum urea and total protein. Serum protein fractions were determined by paper electrophoresis using LKB paper electrophoresis equipment (LKB buffer pH 8.6, μ = 0.1). Urine examinations and bromsulphthalein (BSP) liver function tests were carried out on the control and top dose groups every four months.
Sacrifice and pathology:
At autopsy, a gross pathological examination was made and the major visceral organs weighed. Histopathological examination was made of haematoxylin and eosin stained sections of liver, thyroid, parathyroid, heart, lungs, spleen, kidneys, adrenals, small and large intestines, stomach, pancreas, skin, skeletal muscle, salivary glands, lymph nodes, bladder, testes, prostate, uterus, ovaries, Fallopian tubes, eye and brain of all animals.
Other examinations:
No data
Statistics:
Not specified

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Details on results:
Throughout the experiment, the general health, behaviour and body weights of the dogs remained unaffected by the feeding of Ionox 330. During the first 12 weeks of the experiment, one female from each treatment group, all from one litter, became lethargic, anorexic and pyrexial. Other symptoms developed and distemper was diagnosed. These symptoms occurred four weeks after vaccination for distemper, leptospirosis and hepatitis of all puppies in the kennel. As a result of this infection, the 2,000 ppm dog was killed in the fourth week and the 10,000 ppm animal in the 12th week of the experiment. The pathological findings confirmed the clinical diagnosis. During the eighth week, the affected control group female recovered within seven days and showed no further effects.
No changes in organ weights or organ/body weight ratios were observed. The haematology and clinical chemistry of the dogs remained unaffected throughout the exposure, the minor variations observed not being considered of any biological significance.
At autopsy, no gross or microscopic changes attributable to the exposure were found. Lesions associated with ascarid infestation were detected in the livers of both treated and control animals. Evidence of pulmonary and renal disease was also found in all the animals.

Effect levels

Dose descriptor:
NOEL
Effect level:
10 000 ppm
Based on:
test mat.
Sex:
male/female

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

Table 1 – Summary of the pathological findings in dogs fed Ionox 330 for two years

1a - Dogs killed during the experiment

Dietary Concn (ppm)

Animal Number

Time of Exposure (weeks)

Gross and microscopic pathological findings

2000

357 Female

4

Focal meningitis, Peri-arteritis of large intestine. Haemorrhage of lymph nodes.

10000

360 Female

12

Bronchopneumonia with numerous abscesses.

1b - After two years

Dietary Concn (ppm)

Animal Number

Sex

Gross and microscopic pathological findings

10000

340

Male

Hepatic granulomas. Nephritis.

 

348

Male

Negative.

 

351

Male

Hepatic granulomas. Nephritis.

 

352

Male

Bronchopneumonia. Pyelonephritis.

 

321

Female

Pneumonitis with granulomas. Hepatic granulomas. Nephritis.

 

343

Female

Hepatic granulomas.

 

356

Female

Hepatic granulomas. Nephritis.

2000

361

Male

Pulmonary fibrosis. Parathyroid adenoma.

 

363

Male

Focal pneumonitis.

 

339

Male

Hepatic granulomas. Nephritis.

 

354

Male

Hepatic granulomas.

 

342

Female

Pneumonitis with granulomas. Hepatic ganulomas. Pyelitis.

 

338

Female

Nephritis and pyelitis.

 

345

Female

Hepatic granulomas. Renal abscess. Resolving pneumonia.

0

347

Male

Hepatic – peripertal inflammation.

 

349

Male

Focal pulmonary fibrosis. Fatty infiltration myocardium.

 

350

Male

Interstitial pneumonitis with fibrosis. Mild pyelitis.

 

353

Male

Bronchitis.

 

355

Male

Patchy pneumonitis. Hepatic granulomas.

 

359

Female

Hepatic granulomas. Nephritis.

 

341

Female

Pneumonitis. Hepatic granulomas. Pyelitis.

 

344

Female

Hepatic granulomas. Nephritis.

 

346

Female

Patchy pneumonitis. Hepatic granulomas. Nephritis.

 

358

Female

Hepatic granulomas.

 

Table 2 – The organ weights and organ/body weight ratios of dogs fed Ionox 330 for two years

Dietary Concn (ppm)

Number of Animals

Terminal body wt (kg)

Organ weight (g)

Brain

Heart

Liver

Kidneys

Testes

Males

 

0

5

14.8

83.5

126.1

451

678

16.2

2000

4

15.2

85.4

128.3

487

71

17.2

10000

4

14.9

84.6

118.6

410

67

17.9

S.E. of a treatment mean

5

± 0.61

± 3.28

± 6.76

± 32.5

± 3.2

± 1.05

4

± 0.69

± 3.67

± 7.55

± 36.3

± 3.6

± 1.05

Females

 

0

5

12.5

78.3

91.7

349

48

 

2000

3

13.0

74.8

93.7

332

46

 

10000

3

13.3

77.3

96.0

349

51

 

S.E. of a treatment mean

5

± 0.42

± 2.62

± 4.73

± 25.2

± 4.2

 

3

± 0.54

± 3.38

± 6.11

± 32.6

± 5.4

 

 

Organ/body weight ration (g/100 g body weight)

Male

 

0

5

 

0.57

0.85

3.02

0.45

0.11

2000

4

 

0.57

0.85

3.21

0.47

0.12

10000

4

 

0.57

0.80

2.75

0.45

0.12

S.E. of a treatment mean

5

 

± 0.033

± 0.044

± 0.145

± 0.018

± 0.011

4

 

± 0.037

± 0.049

± 0.162

± 0.020

± 0.011

Females

 

0

5

 

0.61

0.73

2.77

0.38

 

2000

3

 

0.59

0.72

2.54

0.35

 

10000

3

 

0.59

0.72

2.63

0.38

 

S.E. of a treatment mean

5

 

± 0.013

± 0.028

± 0.135

± 0.021

 

3

 

± 0.016

± 0.036

± 0.174

± 0.027

 

 

Applicant's summary and conclusion

Conclusions:
In conclusion, this experiment shows Ionox 330 produces no effects when fed to dogs for two years at dietary concentrations of up to 10,000 ppm.
Executive summary:

lonox 330 was fed to dogs for two years at dietary concentrations of 0, 2,000 and 10,000 ppm. Observations were made on the health, behaviour and body weight of all animals. Blood samples were taken regularly for routine haematologica1 and clinical chemical examinations. Liver function tests were carried out at intervals on the control and top dose groups. At autopsy, major visceral organs were weighed and a histopathological examination made of all tissues.

The general health, behaviour and growth rate of the dogs remained unaffected throughout the experiment. No changes were observed in the organ weights and organ/body weight ratios, haematology and clinical chemistry of the animals.No lesion attributable to the exposure to lonox 330 was detected.

A dietary concentration of 10,000 ppm lonox 330 did not produce any toxicological effects when fed to dogs for two years.